LOCUS Exported 7045 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION mammalian expression of ARAF S214C. ACCESSION . VERSION . KEYWORDS pBABEpuro-ARAF_S214C SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7045) TITLE pBABEpuro-ARAF JOURNAL Unpublished REFERENCE 2 (bases 1 to 7045) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7045 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 8..477 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 542..741 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 934..1350 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1318..1334 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" protein_bind 1378..1402 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(3224..3248) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(3284..3304) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3289..3618 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3469..3604 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3531..3550 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3628..4227 /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" primer_bind complement(3628..3647) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4124..4144 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 4326..4919 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(5120..5137) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5291..5879) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5371..5390) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6057..6917) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6680..6699 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6918..7022) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc 361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg 541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta 601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa 661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc 721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg 781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt 841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt 901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt 961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc 1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg 1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca 1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct 1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt 1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt 1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc cagtgtggtg gtacatcaca 1381 agtttgtaca aaaaagttgg catggagcca ccacggggcc cccctgccaa tggggccgag 1441 ccatcccggg cagtgggcac cgtcaaagta tacctgccca acaagcaacg cacggtggtg 1501 actgtccggg atggcatgag tgtctacgac tctctagaca aggccctgaa ggtgcggggt 1561 ctaaatcagg actgctgtgt ggtctaccga ctcatcaagg gacgaaagac ggtcactgcc 1621 tgggacacag ccattgctcc cctggatggc gaggagctca ttgtcgaggt ccttgaagat 1681 gtcccgctga ccatgcacaa ttttgtacgg aagaccttct tcagcctggc gttctgtgac 1741 ttctgcctta agtttctgtt ccatggcttc cgttgccaaa cctgtggcta caagttccac 1801 cagcattgtt cctccaaggt ccccacagtc tgtgttgaca tgagtaccaa ccgccaacag 1861 ttctaccaca gtgtccagga tttgtccgga ggctccagac agcatgaggc tccctcgaac 1921 cgccccctga atgagttgct aaccccccag ggtcccagcc cccgcaccca gcactgtgac 1981 ccggagcact tccccttccc tgccccagcc aatgcccccc tacagcgcat ccgctccacg 2041 tgcactccca acgtccatat ggtcagcacc acggccccca tggactccaa cctcatccag 2101 ctcactggcc agagtttcag cactgatgct gccggtagta gaggaggtag tgatggaacc 2161 ccccggggga gccccagccc agccagcgtg tcctcgggga ggaagtcccc acattccaag 2221 tcaccagcag agcagcgcga gcggaagtcc ttggccgatg acaagaagaa agtgaagaac 2281 ctggggtacc gggactcagg ctattactgg gaggtaccac ccagtgaggt gcagctgctg 2341 aagaggatcg ggacgggctc gtttggcacc gtgtttcgag ggcggtggca tggcgatgtg 2401 gccgtgaagg tgctcaaggt gtcccagccc acagctgagc aggcccaggc tttcaagaat 2461 gagatgcagg tgctcaggaa gacgcgacat gtcaacatct tgctgtttat gggcttcatg 2521 acccggccgg gatttgccat catcacacag tggtgtgagg gctccagcct ctaccatcac 2581 ctgcatgtgg ccgacacacg cttcgacatg gtccagctca tcgacgtggc ccggcagact 2641 gcccagggca tggactacct ccatgccaag aacatcatcc accgagatct caagtctaac 2701 aacatcttcc tacatgaggg gctcacggtg aagatcggtg actttggctt ggccacagtg 2761 aagactcgat ggagcggggc ccagcccttg gagcagccct caggatctgt gctgtggatg 2821 gcagctgagg tgatccgtat gcaggacccg aacccctaca gcttccagtc agacgtctat 2881 gcctacgggg ttgtgctcta cgagcttatg actggctcac tgccttacag ccacattggc 2941 tgccgtgacc agattatctt tatggtgggc cgtggctatc tgtccccgga cctcagcaaa 3001 atctccagca actgccccaa ggccatgcgg cgcctgctgt ctgactgcct caagttccag 3061 cgggaggagc ggcccctctt cccccagatc ctggccacaa ttgagctgct gcaacggtca 3121 ctccccaaga ttgagcggag tgcctcggaa ccctccttgc accgcaccca ggccgatgag 3181 ttgcctgcct gcctactcag cgcagcccgc cttgtgcctt agtgaccaac tttcttgtac 3241 aaagtggtga tgtaggaatt cgccagcaca gtggtcgacc tgtggaatgt gtgtcagtta 3301 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3361 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3421 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3481 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3541 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3601 ggcctaggct tttgcaaaaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc 3661 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc 3721 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa 3781 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc 3841 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc 3901 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc 3961 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg 4021 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc 4081 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac 4141 gagcggctcg gcttcaccgt caccgccgac gtcgagtgcc cgaaggaccg cgcgacctgg 4201 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa 4261 aggagcgcac gaccccatgc atcgataaaa taaaagattt tatttagtct ccagaaaaag 4321 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc 4381 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat 4441 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 4501 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 4561 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 4621 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 4681 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 4741 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc 4801 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 4861 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 4921 ccgacttgtg gtctcgctgc cttgggaggg tctcctctga gtgattgact acccgtcagc 4981 gggggtcttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac 5041 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 5101 attggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 5161 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 5221 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 5281 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 5341 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 5401 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 5461 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 5521 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 5581 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 5641 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 5701 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 5761 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 5821 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5881 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6001 gaagtttgcg gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 6061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 6121 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 6181 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 6241 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 6301 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 6361 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 6421 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 6481 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 6541 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 6601 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 6661 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 6721 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 6781 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 6841 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 6901 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 6961 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 7021 cgcacatttc cccgaaaagt gccac //